LOCUS       sequence-257509-        5667 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5667)
  AUTHORS   Galvez-Santisteban M, Rodriguez-Fraticelli AE, Bryant DM, 
            Vergarajauregui S, Yasuda T, Banon-Rodriguez I, Bernascone I, Datta 
            A, Spivak N, Young K, Slim CL, Brakeman PR, Fukuda M, Mostov KE, 
            Martin-Belmonte F
  TITLE     Synaptotagmin-like proteins control the formation of a single apical
            membrane domain in epithelial cells.
  JOURNAL   Nat Cell Biol. 2012 Jul 22. doi: 10.1038/ncb2541.
  PUBMED    22820376
REFERENCE   2  (bases 1 to 5667)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat Cell 
            Biol. 2012 Jul 22. doi: 10.1038/ncb2541."
COMMENT     Sequence Label: pEGFP-C1 hSlp4-a C2AB
FEATURES             Location/Qualifiers
     source          1..5667
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        114..417
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        418..621
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     572..592
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      660..669
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(666..668,669..671,672..1382)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(666..668,669..671,672..1382)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(711..732)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(972..991)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1319..1340
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2508..2629
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2545..2564)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2599..2618
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2636..3091)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2773..2794)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2985..3004
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3118..3222
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3224..3581
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3224..3415
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3247..3267)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3432..3567
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3616..4410
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3616..4410
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3670..3689)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4280..4299
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4598..4617)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4642..4689
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5018..5606
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5507..5526
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat
       61 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
      121 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
      181 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
      241 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
      301 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
      361 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
      421 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
      481 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
      541 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
      601 gaggtctata taagcagagc tggtttagtg aaccgtcaga tccgctagcg ctaccggtcg
      661 ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
      721 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
      781 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
      841 ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca
      901 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
      961 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     1021 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     1081 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
     1141 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
     1201 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     1261 accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
     1321 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     1381 agtccggact cagatctcga gctcaagctt cgaattctaa catctttgtg actggcagga
     1441 ttgccttttc cctgaagtat gagcagcaaa cccagagtct ggttgtccat gtgaaggagt
     1501 gccatcagct ggcctatgct gatgaagcca agaagcgctc taacccatat gtgaagactt
     1561 accttctgcc tgacaagtcc cgccaaggaa aaagaaaaac cagcatcaag cgggacactg
     1621 ttaatccact atatgatgag acgctgaggt atgagatccc agaatctctc ctggcccaga
     1681 ggaccctgca gttctcagtt tggcatcatg gtcgttttgg cagaaacact ttccttggag
     1741 aggcagagat ccagatggat tcctggaagc ttgataagaa actggatcat tgcctccctt
     1801 tacatggaaa gatcagtgct gagtccccga ctggcttgcc atcacacaaa ggcgagttgg
     1861 tggtttcatt gaaatacatc ccagcctcca aaacccctgt tggaggtgac cggaaaaaga
     1921 gtaaaggtgg ggaaggggga gagctccagg tgtggatcaa agaagccaag aacttgacgg
     1981 ctgccaaagc aggagggact tcagacagct ttgtcaaggg atacctcctt cccatgagga
     2041 acaaggccag taaacgtaaa actcctgtga tgaagaagac cctgaatcct cactacaacc
     2101 atacatttgt ctacaatggt gtgaggctgg aagatctaca gcatatgtgc ctggaactga
     2161 ctgtgtggga ccgggagccc ctggccagca atgacttcct gggaggggtc aggctgggtg
     2221 ttggcactgg gatcagtaat ggggaagtgg tggactggat ggactcgact ggggaagaag
     2281 tgagcctgtg gcagaagatg cgacagtacc cagggtcttg ggcagaaggg actctgcagc
     2341 tccgttcctc aatggccaag cagaagctgg gtttatgagg atccaccgga tctagataac
     2401 tgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     2461 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     2521 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     2581 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa cgcgtaaatt
     2641 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     2701 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     2761 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     2821 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     2881 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     2941 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     3001 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     3061 gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc
     3121 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     3181 taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc
     3241 agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3301 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     3361 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     3421 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     3481 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     3541 agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga
     3601 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     3661 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     3721 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     3781 gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     3841 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     3901 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     3961 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     4021 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     4081 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     4141 gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     4201 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     4261 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     4321 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     4381 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     4441 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     4501 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     4561 tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac
     4621 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt
     4681 cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga
     4741 gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt
     4801 cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag
     4861 gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     4921 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     4981 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     5041 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     5101 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     5161 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     5221 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     5281 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     5341 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     5401 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     5461 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     5521 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     5581 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     5641 ccttttgctg gccttttgct cacatgt
//
